LOCUS       sequence-323490-        6099 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Acute Depletion of BRG1-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6099)
  AUTHORS   Zhu Y, Denholtz M, Lu H, Murre C
  TITLE     Calcium signaling instructs NIPBL recruitment at active enhancers 
            and promoters via distinct mechanisms to reconstruct genome 
            compartmentalization.
  JOURNAL   Genes Dev. 2021 Jan 1;35(1-2):65-81. doi: 10.1101/gad.343475.120. 
            Epub 2020 Dec 17.
  PUBMED    33334824
REFERENCE   2  (bases 1 to 6099)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            Dev."; date: "2021-01-1"; volume: "35(1-2):65-81. doi"; pages: " 
            10.1101/gad.343475.120. Epub 2020 Dec 17"
COMMENT     Sequence Label: pFlox_N-BRG1_BSD_FKBPF36V_EYFP
FEATURES             Location/Qualifiers
     source          1..6099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             876..1271
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1281..1337
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1338..1361
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1371..1691
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1698..1724
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1725..1727,1728..2438)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1767..1788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2028..2047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2375..2396
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3602..3620)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3641..3657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3641..3657)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3654..3676)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3665..3681
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3689..3695,3696..3713,3714..3719))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3734..3755
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3872..3889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4043..4631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4123..4142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4802..5593,5594..5662))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5425..5444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5663..5767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5835..5853
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5891..5913)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6013..6032
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccgcggc ggccgcacta gtggccgcgg
      181 cggccgcact agtggccgcg gcggccgcag cttgcctgca ggtcgacgat tagcactcac
      241 tgattgtttt tgtgtgtctg tctgactgcc tgtcttgttc atgaccttca tgaacaagag
      301 aacagatgac tccacagtta aaggataatt tgagagttgc cccacagtct gctggggacc
      361 tagttgatct tggaggtaat gggaggacat tgtcccactg accacatcaa gtgtagcttc
      421 ctagagtgat tattggtcag tttccccagt ctgtcacttt atagttagat aaggctgggc
      481 aatacgcaga agtaattttg tggtagggtg gggagtgaga cagggtctcc ctatgcagtc
      541 ctggctggcc tggaactagt atagaactca aattaagaga tcctctgcct ccctatttct
      601 ggggtcatgg acatgcacca tcattccctg ctttctgagc tgatttcttt tcctgagata
      661 acattgtttt tcccaaaaag aataaaagaa aggtttggaa caaaggttag gtatcaggcc
      721 tacaagatgg ggctcagtgg tctgggtgaa cctgactgac ctttgggccc ttctttgctg
      781 ttcttgacat aactgaaccc ctgtcccaga ttctaggatg accagaattt ctttccaaca
      841 ggagaccagt gcctgtagct ccagtaaagg tcgacatggc caagcctttg tctcaagaag
      901 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
      961 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     1021 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     1081 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     1141 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     1201 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     1261 tgtgggaggg cggttccgga gccacgaact tctctctgtt aaagcaagca ggagacgtgg
     1321 aagaaaaccc cggtcccgat tacaaggatg acgatgacaa gggttccggc ggcgtgcagg
     1381 tggagactat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     1441 tgcactacac cgggatgctt gaagatggaa agaaagttga ttcctcccgg gacagaaaca
     1501 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     1561 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     1621 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     1681 taaaactgga agtcgattat ccgtacgacg taccagacta cgcagtgagc aagggcgagg
     1741 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1801 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1861 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     1921 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1981 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2041 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2101 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2161 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2221 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2281 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     2341 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2401 ccgccgggat cactctcggc atggacgagc tgtacaaggg tggaggtgga tctggtggcg
     2461 gcggttcagg taccttgggt gggactcctc ggcctggtcc ttccccaggc cctggtcctt
     2521 cacctggtgc aatgctgggt cctagccctg gcccctcacc aggttctgcc cacagcatga
     2581 tggggccaag cccaggacct ccttcagcag gacatcccat gcccacccag gggcctggag
     2641 ggtaccccca ggacaacatg catcagatgc acaaggtagg catcctgtga tctcagtcct
     2701 gtactctgtt cctgagactt gccttggtgt ctcatacaaa gctgccagag cctccggaca
     2761 ttagaggaga gaggtctgtg gaggtgactt ggcttagttc agggtcctac cctaaagtgt
     2821 gcttctttga acaaaggaat caggctgttg tcctgagcca ggaggctgag cagccatttc
     2881 actctacgtt tcccattctc cctatgcttt cttttgttaa aaggaattcc cagattcctc
     2941 taaggacgat cctctggctg cactggtggc ctacatactt gaagcagtgg ggagtattga
     3001 gtctgcttct gagcttgtac cccagactgc catcttcctt tggtaaatgt gtcagcattg
     3061 cttttgccag caaggctgcc aggcagcaat cctgtcctta ggccttggtg ttcctcttag
     3121 gactggcgca gtagaatgtc agcatttatc tgggctgacc cacagaggct ggggtgtaag
     3181 aagatattgg aggctgcagg agggttcacc atagtggcat tgtcagttgc tcctcttccc
     3241 tgcagggttg tcctctgttt gcttcctaat cctgcaagta taggttgtgt tccctatcca
     3301 caaatgagga aggccctcac atgcctgtct cattcctcgg cctggcttgt ccttgtactc
     3361 tgcttccaga catctccatc tggctcccca gttctggatg gttgtgtcat cttagtagag
     3421 tacttatctt gctctttccc ctatgaagga agctctgggt cccatgtagg agcccaccgt
     3481 ctggtttaca gggccctcca tgtccggtaa gctgcatctc ggtaccctcg aggtcgatcg
     3541 acggtatcga taagcttgat ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     3601 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3661 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3721 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3781 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3841 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3901 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3961 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4441 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4561 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4681 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4741 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4801 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4861 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4921 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4981 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5041 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5101 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5161 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5221 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5281 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5341 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5401 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5461 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5521 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5581 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5641 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5701 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5761 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5821 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     5881 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5941 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6061 taccgcacag atgcgtaagg agaaaatacc gcatcaggc
//
